A COS-cell-based system for rapid production and quantification of scFv::IgCκ antibody fragments

Single-chain Fv (scFv) antibody'(Ab) fragments were transiently produced in COS-1 cells utilizing a mammalian expression vector featuring a murine immunoglobulin (Ig) light-chain leader sequence for efficient secretion and a murine Ig kappa constant domain (IgCκ) for detection. Several hundred...

Ausführliche Beschreibung

Gespeichert in:

Bibliographische Detailangaben
Veröffentlicht in:	Gene 1995-12, Vol.166 (2), p.273-276
Hauptverfasser:	Ridder, Rüdiger, Geisse, Sabine, Kleuser, Beate, Kawalleck, Petra, Gram, Hermann
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals Base Sequence Cell Line Cercopithecus aethiops chimeric single-chain antibody fragment Cloning, Molecular - methods DNA Primers - chemistry Gene Expression Genes, Immunoglobulin - genetics Immunoglobulin kappa-Chains - genetics Immunoglobulin Variable Region - genetics Mammalian expression Molecular Sequence Data murine IgCκ tag rabbit monoclonal antibody Recombinant Fusion Proteins
Online-Zugang:	Volltext
Tags:	Tag hinzufügen Keine Tags, Fügen Sie den ersten Tag hinzu!

Beschreibung
Zusammenfassung:	Single-chain Fv (scFv) antibody'(Ab) fragments were transiently produced in COS-1 cells utilizing a mammalian expression vector featuring a murine immunoglobulin (Ig) light-chain leader sequence for efficient secretion and a murine Ig kappa constant domain (IgCκ) for detection. Several hundred milliliters of supernatants from large-scale COS cell transfections were sufficient to purify the scFv::IgCK fusion proteins by one-step affinity chromatography utilizing an immobilized rat anti-mouse IgCκ monoclonal Ab. Furthermore, the murine IgCκ domain allowed for accurate quantification of the scFv::IgCκ fusion protein secreted into the COS cell supernatant by a sandwich enzyme-linked immunosorbent assay (S-ELISA).
ISSN:	0378-1119 1879-0038
DOI:	10.1016/0378-1119(95)00638-9